Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (29): 5355-5360.doi: 10.3969/j.issn.2095-4344.2012.29.008

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Histological structure of porcine aortic valve scaffolds decellularized by using different methods

Ma Hao1, Wang Qi1, Shi Hai-yan1, Wang Li-xin1, Zhang Xiao1, Li Ze-jian2, Miao Qi2   

  1. 1Department of Cardiovascular Surgery, General Hospital of Chinese People’s Armed Police Forces, Beijing 100039, China;
    2Department of Cardiothoracic Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100730, China
  • Received:2011-12-24 Revised:2011-12-24 Online:2012-07-15 Published:2012-07-15
  • Contact: Wang Qi, Doctor, Professor, Master’s supervisor, Department of Cardiovascular Surgery, General Hospital of Chinese People’s Armed Police Forces, Beijing 100039, China wangqi1856@sina.com
  • About author:Ma Hao☆, Doctor, Attending physician, Department of Cardiovascular Surgery, General Hospital of Chinese People’s Armed Police Forces, Beijing 100039, China mc_volunteer@163.com

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Abstract:

BACKGROUND: An ideal decellularization method cannot only remove donorcells completely to decrease immunogenicity, but also reserve the extracellular matrix of natural valve, such as collagen and elastic fibers to keep sufficient mechanical strength.
OBJECTIVE: To prepare porcine aortic valve scaffolds decellularized by different methods and to compare their tissue structures, as well as to explore the most effective methods for preparing decellularized valve scaffolds.
METHODS: Totally 20 fresh porcine aortic valve scaffolds were randomly divided into four groups: control group, detergent group, enzymatic digestion group and detergent+enzymatic group. Decellularized valve scaffolds in the latter three groups were prepared by Triton X-100, Trypsin and Triton X-100+Trypsin methods, respectively. The valve scaffolds were contrasted by gross observation, hematoxylin-eosin and Mallory-Heidenhain staining, scanning and transmission electron microscope.
RESULTS AND CONCLUSION: (1) After decellularized, the valve leaflets in the detergent group were soft and smooth. Besides, Hematoxylin-eosin staining showed that the cells were removed from the valve, but few nucleoli remained and fibers were arranged regularly. Mallory-Heidenhain staining showed that the blue collagen and the red elastin were interlaced. Electron microscope observation showed that the fibers presented with wave-like arrangement and lateral stripes could be observed. (2)The valve leaflets in the enzymatic digestion group were collapsed partly, but their endothelium was smooth. Hematoxylin-eosin staining showed that the cells were removed completely and the connective tissue structures were loose. Mallory-Heidenhain staining showed that collagen and elastin were found reticulated interlacing. Electron microscope showed that some fibers were ruptured, but lateral stripe existed. (3)The valve leaflets in detergent + enzymatic group was collapsed partly, but their endothelium was smooth. Hematoxylin- eosin staining showed that the cells were removed completely from valve. Mallory-Heidenhain staining showed that collagen and elastin showed parallel arrangement. Electron microscope observation showed that fibers were intact but scattered, and lateral stripe can be observed. These findings suggest that Triton X-100, Trypsin and Triton X-100+ Trypsin methods can remove valve cells effectively and keep the fibers intact; moreover, the Triton X-100 + Trypsin method is more effective.

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